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Lectin binding to olfactory system in a shark, Scyliorhinus canicula.

Lectin histochemical studies were performed on the olfactory system of Scyliorhinus canicula to identify specific glycoconjugates on the cell surface of primary olfactory neurons. The olfactory receptor cells, the olfactory nerve fibers and their terminals in the bulbs were labelled with SBA, BSA-I and BSA-I-B4. The lectin staining patterns indicate that the membranes of small-spotted catshark olfactory neurons had glycoproteins with alpha-galactose residues. This carbohydrate moiety could be related to modulation of the cell-cell interactions in the olfactory system.

Animals↗

Neurogenesis in the olfactory bulb of adult zebrafish.

Cell genesis in the adult brain of zebrafish, with specific reference to the olfactory bulbs, was examined using bromodeoxyuridine immunocytochemistry. Mature fish were exposed to a 1% solution of the thymidine analog 5-bromo-2'-deoxyuridine for 1 h and then killed after short (4-h) or long (3-4-week) survival periods. A monoclonal antibody to bromodeoxyuridine allowed visualization of cells that incorporated the drug during the S phase of mitosis. Four hours after administration of the drug, antibody-labeled cells were found almost exclusively in the proliferative zones around the ventricles and in the cerebellum. Very few labeled nuclei were seen in other locations in the brain, indicating that cell genesis occurs in discrete regions in adults. The few labeled profiles in the olfactory bulbs were located in the olfactory nerve layer; these profiles had the morphology of glial nuclei and did not stain with a neuronal marker, the Hu antibody. After longer survival times, labeled cells were present throughout the layers of the olfactory bulb, and many of the immunoreactive profiles in the internal cell layer were also labeled with the Hu antibody, indicating that they are likely adult-formed interneurons. Thus, neurogenesis continues in the olfactory bulb of adult zebrafish. Understanding the process of the generation of new neurons in the brain of adult animals can lead to important insights into neural regeneration and adult plasticity.

Animals↗

Ketamine and MK801 attenuate paired pulse inhibition in the olfactory bulb of the rat.

We have investigated the effects of the phencyclidine like-compounds ketamine and MK801 on the evoked field potentials of rat olfactory bulb. Low doses of ketamine (3-6 mg/kg) blocked the inhibition of mitral cells by granule cells evoked by stimulation of lateral olfactory tract fibres or by stimulation of olfactory nerve. This blockade was not accompanied by a decrease in granule cell excitation as revealed by field potential recording. MK801 had a similar effect on the inhibition of mitral cells evoked by stimulation of the lateral olfactory tract. As ketamine does not influence the inhibitory action of GABA (Anis et al. 1983) these results suggest that both ketamine and MK801 block inhibition by an action on intrinsic excitatory feed-back circuits in the olfactory bulb.

Action Potentials↗

Acetylcholinesterase activity in the developing olfactory bulb: a biochemical study on normal maturation and the influence of peripheral and central connections.

In the newborn rat olfactory bulb (OB), the specific activity of acetylcholinesterase (AChE) is about 20% of the adult value. During postnatal development, the specific activity remains unchanged until day 10; a growth spurt of 5X is observed between days 10 and 25, when adult values (90 nmoles ACh/min/mg protein; 6 nmol/min/mg wet wt.) are reached. However, total activity shows continuous increase slowly at first and rapidly between days 10 to 30, reaching a plateau by day 60. Between birth to day 60 total activity increases 65X. To determine the influence of peripheral and central connections on the development of AChE activity in the OB, rats were subjected to unilateral olfactory denervation and/or transection of olfactory peduncle, carried out either neonatally or at day 30; the bulbs were assayed a month later (days 30 and 60 respectively). It was found that in both neonatal and 30-day-old rats, denervation caused a 15% decrease in total activity, while transection led to more than 60% reduction. In the older rats, the reduction due to transection represented the degenerative loss of activity, but in the neonatally transected bulbs the growth of some cholinergic elements continued although very slowly. Between birth to day 30, total AChE activity increased only 12X in completely isolated OB, 25X in transected OB and 40X in denervated OB, compared to 45-50X in control OB. In the transected and isolated bulbs specific activity of AChE was also reduced significantly (25-50% depending on age and operation). These results suggest that while centrifugal fibers are the main source of cholinergic activity in the mature as well as the developing OB, the olfactory nerve and some intrabulbar sources such as cholinergic cells or cholinoceptive membranes also contribute to AChE activity in the OB. These intrinsic sources of AChE activity can persist and even show some growth in the developing olfactory bulb in the absence of the centrifugal fibers and/or the olfactory afferents.

Acetylcholinesterase↗

[Subfrontal schwannoma: case report].

Schwannoma are benign, slowly growing nerve sheath tumors. They can arise from any peripheral nerve containing Schwann cells including distal portions of cranial nerves. Intracranial schwannomas arising from a subfrontal lesion are very rare. We report a case of subfrontal schwannoma in a 38-year-old woman who presented with seizures in February 2003. MRI features resembled an olfactory groove meningioma. The left carotid angiography revealed that the tumor was supplied from the anterior ethomoidal artery, but no remarkable tumor stain was detected. Bifrontal craniotomy with total excision of the tumor revealed a tumor in the left subfrontal lesion. The tumor was attached to the falx, the left frontal lobe, and the skull base, but those structures were not invaded. Unfortunately, we could not detect the left olfactory nerve. Subsequent histological examination proved that this tumor was schwannoma. A review of the literature found 27 cases including this case previously described. Arguments are raised that schwannomas in a subfrontal lesion are comparable to or even occur as intracranial schwannomas with frontal localization. Subfrontal schwannomas are very rare. For this reason they have not been diagnosed preoperatively. Further cases of subfrontal schwannoma are necessary to solve the enigma of the origin of this tumor.

Adult↗

Lectin histochemical study of olfactory neurons in the eel.

Lectin histochemical studies were performed on paraffin embedded sections of the olfactory system of the eel to identify specific glycoconjugates on the surface of primary olfactory neurons. The olfactory receptors, the olfactory nerve fibres and their terminals in the bulbs were labelled with the lectins (SBA, BSA-I, BSA-I-B4 and DBA) HRP-conjugated or biotinylated. The lectin staining patterns indicate that the membrane of olfactory neurons of the eel had oligosaccharides with alpha-galactose and alpha-N-acetyl-D-galactosamine residues. These findings represent the demonstration of a molecular probe that recognizes specific sets of neurons. The identical histochemical features previously described in the olfactory neurons in amphibians suggest that these carbohydrate moieties might to related to modulation of the cell-cell interactions in the olfactory system of vertebrates.

Anguilla↗

Axonal projections and conduction properties of olfactory peduncle neurons in the rat.

Electrophysiological methods were employed to study the axonal properties of the neurons of anterior olfactory nucleus (AON), transition zone (TZ), and rostral prepyriform cortex (RPPC) and their projections towards the ipsilateral and contralateral olfactory bulb (IOB, COB) in the rat. Of 91 antidromically driven cells, 39 (43%) and 32 (35%) responded to IOB and COB stimulation, respectively; 20 (22%) were discharged from both bulbs. Collison tests performed on the latter group indicated that these neurons have a short main axon which divides near the soma, projecting one branch to the COB and a thinner one toward the IOB. Mean conduction velocities of axons projecting to the IOB and the COB were 0.4 m/s and 0.7 o/s, respectively, the faster conducting axons having shorter refractory periods. Of the 38 neurons tested, 92% showed decreases in threshold and latency (up to 20% of control antidromic latency) after a test volley that was preceded by a conditioning pulse at intervals of 20--215 ms. Latency decreases were greater for slowly conducting axons than for the faster ones. These after-effects of impulse activity in OB afferent axons were attributed to the presence of a supernormal period of increased conduction velocity and excitability similar to that found in the olfactory nerve (Bliss and Rosenberg, 1974).

Animals↗

The effect of intranasal zinc sulfate treatment on odor-mediated behavior and on odor-induced metabolic activity in the olfactory bulbs of neonatal rats.

This study describes the effect of lesions of the peripheral olfactory receptor sheet on odor-mediated behavior and on odor-induced metabolic activity in the neonatal olfactory bulb. Nine-day-old rats were treated by intranasal irrigation with a solution of 1% ZnSO4 or 5% ZnSO4 or saline. At 1 and 5 days following treatment they were tested for maternally-directed behaviors which are mediated in large part by olfactory cues. At 1 day following treatment the pups treated with ZnSO4 solutions had significant deficits in their odor-directed behavior. By 5 days however, most pups treated with 1% ZnSO4 had recovered to control levels, while the pups treated with 5% ZnSO4 continued to exhibit profound behavioral deficits. Following the behavioral testing, the pups were tested using the 2-deoxyglucose method for amyl acetate odor-induced metabolic activity in their olfactory bulbs. Control pups had large regions of high focal activity in their anterolateral and caudomedial olfactory bulbs. The 1% ZnSO4 pups had smaller foci which were confined primarily to the medial olfactory bulb. The 5% ZnSO4 pups had very little focal uptake in their olfactory bulbs. There was a clear correlation between intact olfactory nerve and glomerular layers and sites of increased functional activity. The results of this study demonstrate that there is a relationship between the degree of behavioral deficit and the extent of odor-induced functional activity in the olfactory bulb.

Animals↗

Silver lampreys (Ichthyomyzon unicuspis) lack a gonadotropin-releasing hormone- and FMRFamide-immunoreactive terminal nerve.

The terminal nerve is a ganglionated cranial nerve with peripheral processes that enter the nasal cavity and centrally directed processes that enter the forebrain. Members of all classes of gnathostomes have been found to possess a terminal nerve, some components of which demonstrate immunoreactivity to the peptides Phe-Met-Arg-Phe-NH2 (FMRFamide) and gonadotropin-releasing hormone (GnRH). To explore the possibility that lampreys possess a terminal nerve, we examined the distribution of these peptides in the silver lamprey, Ichthyomyzon unicuspis, by using antisera to FMRFamide and to four forms of GnRH. We found cells with FMRFamide-like immunoreactivity in the preoptic area and the isthmal gray region of the mesencephalon, and found labeled fibers throughout the preoptic-infundibular region. Occasional labeled fibers were scattered through many regions of the brain, including the optic nerve and olfactory bulb; however, unlike species that possess a terminal nerve, lampreys have no immunoreactive cells or fibers in the olfactory nerve or nasal epithelia. In addition, we observed GnRH-immunoreactive cell bodies in the preoptic area of all animals and in the ventral hypothalamus of one individual. Most of the labeled fibers extended ventrally to the hypothalamus, with other fibers extending throughout the striatum and hypothalamic-neurohypophyseal region. A few fibers in other regions, including the optic nerve, were also labeled; we detected no immunoreactivity in the olfactory bulb, olfactory nerve, or nasal epithelia. The use of different GnRH antisera resulted in remarkably similar patterns of labeling of both cells and fibers. In summary, we did not observe either GnRH or FMRFamide-like immunoreactivity in the olfactory regions that represent the typical path of terminal nerve fibers, nor were we able to locate a terminal nerve ganglion. We conclude that lampreys may lack a terminal nerve, and that the previously described fiber bundle extending from the nasal sac to the ventral forebrain may constitute an extra-bulbar olfactory pathway.

Animals↗

Influence of GABA, baclofen and bicuculline on the extracellular K+ activity in the olfactory bulb of the developing rat.

gamma-Aminobutyric acid (GABA) and baclofen were iontophoresed into the glomerular and plexiform layers of the olfactory bulb of rats at different ages. Subsequent changes in K+ activity and mitral cell firing rates were monitored to specify GABA receptor activation. Only bicuculline-insensitive receptors are shown to be already present in the glomerular layer of the newborn rat. The possible distribution and maturation of GABA receptors on the olfactory nerve endings or on the dendrites of the mitral or periglomerular cells is discussed.

Action Potentials↗

Functional heterogeneity of periglomerular cells in the rat olfactory bulb.

The periglomerular cells of the rat olfactory bulb, a virtually unknown population of interneurons, have been studied applying the whole-cell patch-clamp technique to thin slices. A prominent result, obtained under current-clamp conditions, is that these cells appear to be functionally heterogeneous, and show distinct excitability profiles. Voltage-clamp analysis allows the identification of the ionic basis of these differences and suggests a division into at least two classes, based on the characteristics of the K+ conductances. The first group displays two K+ conductances (delayed rectifier, gKV, and fast transient, gA) of similar amplitude, and under current-clamp conditions shows the usual outward rectifying behaviour at depolarized potentials. The second group has a large gA, and a small or absent gKV. Consequently, following sustained depolarizations under current-clamp conditions leading to inactivation of gA, these neurons do not show any sign of outward rectification and behave as ohmic elements, as normally observed only at hyperpolarized potentials. The transition ion zinc (10-300 microM) affects gA but not gKV The inactivation process (steady-state curve and rate constant) is strongly altered by Zn2+, the activation process less so; open-channel conductance is not affected. The Zn2+ effect is unlikely to be due to surface charge screening or to a mechanism involving channel block. In view of the substantial presence of zinc ions in the olfactory nerve terminals, its actions on the A-current could be of some relevance for physiological function.

Animals↗

MRI detection of olfactory bulb and tract.

Thirty healthy volunteers underwent MRI with 3D MP-RAGE, 3D CISS and 2D turbo spin echo sequences to compare the detectability of olfactory fibers, bulb, tract, and sulcus. The overall detectability was slightly better using MP-RAGE compared with CISS. Both 3D sequences were superior to 2D turbo spin echo. We therefore recommend including the MP-RAGE sequence in an MR imaging protocol of the olfactory nerve.

Adolescent↗

Golgi, electron-microscopic and combined Golgi-electron-microscopic studies of the mitral cells in the goldfish olfactory bulb.

The local neuronal circuitry of goldfish olfactory bulb was analyzed in Golgi preparations combining light- and electron-microscopy, as well as in routinely prepared ultrastructural preparations. Mitral cells were identified with the light-microscope in Golgi-impregnated thick sections according to the following criteria: (1) cell bodies were distributed irregularly in a wide layer between 100 and 200 micrometer from the surface, (2) cell bodies were larger than other neurons (10-20 micrometer in diameter), and (3) the dendrites were directed toward the superficially-located olfactory nerve layer where they ended as highly branched glomerular tufts. These impregnated cells were examined by electron-microscopy in serial section. The results demonstrate synaptic organization in relation to the mitral cells. (1) Glomerular tufts received afferent input from primary olfactory axons which made Gray's Type I synaptic contacts. These dendrites also had reciprocal dendrodendritic synapses with dendrites of certain non-mitral cells. (2) Dendritic shafts of mitral cells made reciprocal dendritic synapses with dendrites of certain non-mitral cells. (3) Cell bodies and their initial axon segments had reciprocal synapses with certain dendrites but occurred infrequently. In reciprocal synapses, the direction of the Gray Type I (asymmetrical) is away from the mitral cell while those with Gray Type II synapses (symmetrical) are toward the mitral cell. Assuming that the type I synapse is excitatory and Type II is inhibitory, these findings explain the electrophysiological demonstration of self-inhibition discharge found in mitral cells.

Animals↗

Localization of choline acetyltransferase and vasoactive intestinal polypeptide-like immunoreactivity in the nervus terminalis of the fetal and neonatal rat.

Ganglia of the nervus terminalis have been shown to contain luteinizing hormone-releasing hormone (LHRH) immunoreactive cells in several mammalian species. These cells are always accompanied by clusters of cells non-immunoreactive to antiserum to LHRH. Using immunocytochemical procedures, we found choline acetyltransferase (ChAT) and vasoactive intestinal polypeptide (VIP) present in cell bodies and in nerve processes throughout the peripheral, intracranial and central projections of the nervus terminalis. In addition, a dense plexus of substance P (SP) immunoreactive fibers was seen in the nasal mucosa surrounding the nasal glandular acini and blood vessels. A number of SP reactive fibers were traced with the olfactory nerves through the cribriform plate of the ethmoid bone and appeared to enter the brain in the area of the central roots of the nervus terminalis.

Animals↗

Nerve growth factor receptor expression in the young and adult rat olfactory system.

Nerve growth factor (NGF) and its receptor (NGFR) are proteins that have a role in the normal development and survival of neurons in the peripheral and central nervous systems. During development, NGF is necessary for outgrowth of axons and establishment of synapses, and NGFR is the transmembrane protein that binds NGF and brings it into the cell. NGF and NGFR expression in the rat olfactory system have been studied previously, and age differences in NGFR are explored further in this study, using immunocytochemistry and immunoelectron microscopy to determine the changes in two different ages: postnatal day 5 and the adult. Dramatic differences were found in the distribution of NGFR immunoreactivity in the olfactory system of each of the two ages studied. Electron microscopy revealed that glial cells were responsible for this immunoreactivity.

Animals↗

NQO1 activity in the main and the accessory olfactory systems correlates with the zonal topography of projection maps.

The mouse olfactory epithelium (OE) is divided into spatial zones, each containing neurons expressing zone-specific subsets of odorant receptor genes. Likewise, the vomeronasal (VN) organ is organized into apical and basal subpopulations of neurons expressing different VN receptor gene families. Axons projecting from the different OE zones and VN subpopulations form synapses within circumscribed regions in the glomerular layer of the olfactory bulb (OB) and accessory olfactory bulb (AOB), respectively. We here show that mature neurons in one defined zone selectively express NADPH:quinone oxidoreductase (NQO1), an enzyme that catalyses reduction of quinones. Immunohistochemistry and in situ hybridization analyses show non-overlapping expression of NQO1 and the Rb8 neural cell adhesion molecule (RNCAM/OCAM) in OE and axon terminals within glomeruli of the OB. In addition, NQO1 immunoreactivity reveals selective, zone-specific axon fasciculation in the olfactory nerve. VN subpopulations do not show complementary patterns of RNCAM and NQO1 immunoreactivity, instead both genes are co-expressed in apical VN neurons that project to the rostral AOB. These results indicate that one division of both the accessory and the main olfactory projection maps are composed of sensory neurons that are specialized to reduce environmental and/or endogenously produced quinones via an NQO1-dependent mechanism. The role of NQO1 in bioactivation of quinoidal drugs also points to a connection between zone-specific NQO1 expression and zone-specific toxicity of certain olfactory toxins.

Animals↗

Development and distribution of gonadotropin-releasing hormone neuronal systems in the frog (Rana esculenta) brain: immunohistochemical analysis.

The ontogenesis of the gonadotropin-releasing hormone (GnRH) neuronal systems was studied in the brain of the frog, Rana esculenta. Attention was also focussed on the differential distribution of molecular forms of GnRH during development. The first GnRH-immunoreactive neurons appear in the mesencephalon of posterior limb-stage tadpoles. These neurons are shown to contain only chicken [His5,Trp7,Tyr8]GnRH (cGnRH-II). Later in development, mammalian [Tyr5,Leu7,Arg8] GnRH (mGnRH)-like peptide-containing neurons appear simultaneously in the terminal nerve as well as in the anterior preoptic area of the telencephalon. Subsequently, only after metamorphosis, mGnRH-containing neurons appear in the medial septal area of the posterior telencephalon. It is here shown that neurons containing the two forms of GnRH are distributed in distinct brain areas during development and in the adult: mGnRH-immunoreactive neurons in the terminal nerve, olfactory bulb, mediobasal telencephalon, medial septal area, anterior preoptic area, ventrolateral thalamus and infundibulum, whereas cGnRH-II neurons are located in the mesencephalon. We hypothesize that the terminal nerve/forebrain-located GnRH neurons express immunohistochemically late in development and originate extracranially migrating centrally, along the terminal nerve, during development, whereas those located in the mesencephalon express earlier and may have an intracranial site of origin.

Animals↗